Sun Y, Elwell J H, Oberley L W
Radiation Research Laboratory, University of Iowa, Iowa City 52242.
Free Radic Res Commun. 1988;5(2):67-75. doi: 10.3109/10715768809066913.
A simple and sensitive method for the simultaneous visualization of glutathione peroxidase and catalase on polyacrylamide gels is described. The procedure included: (1) running samples on a 7.5% polyacrylamide gel, (2) soaking the gel in a certain concentration of reduced glutathione (0.25-2.0 mM), (3) soaking the gel in GSH plus H2O2 or cumene hydroperoxide, (4) finally staining with a 1% ferric chloride 1% potassium ferricyanide solution. The best concentration of glutathione for simultaneous visualization of glutathione peroxidase in mouse liver homogenates and also it is specific for glutathione peroxidase since other peroxidases such as lactoperoxidase, horseradish peroxidase and glutathione S-transferase cannot be visualized. Using this method, it was found that unlike catalase, glutathione peroxidase is heat resistant (68 degrees C, 1 min), but sensitive to 10 mM sodium iodoacetate.
本文描述了一种在聚丙烯酰胺凝胶上同时可视化谷胱甘肽过氧化物酶和过氧化氢酶的简单且灵敏的方法。该步骤包括：(1) 在7.5%的聚丙烯酰胺凝胶上运行样品，(2) 将凝胶浸泡在一定浓度的还原型谷胱甘肽（0.25 - 2.0 mM）中，(3) 将凝胶浸泡在谷胱甘肽加过氧化氢或异丙苯过氧化氢中，(4) 最后用1%的氯化铁和1%的铁氰化钾溶液染色。用于同时可视化小鼠肝脏匀浆中谷胱甘肽过氧化物酶的最佳谷胱甘肽浓度，并且它对谷胱甘肽过氧化物酶具有特异性，因为其他过氧化物酶如乳过氧化物酶、辣根过氧化物酶和谷胱甘肽S - 转移酶无法可视化。使用该方法发现，与过氧化氢酶不同，谷胱甘肽过氧化物酶耐热（68摄氏度，1分钟），但对10 mM碘乙酸钠敏感。
